Juvenile Hormone Mimic Research Articles

Juvenile hormone suppresses sensory organ precursor determination to block Drosophila adult abdomen morphogenesis

Juvenile hormone (JH) has a classic "status quo" action at both the pupal and adult molts when administrated exogenously. In Drosophila, treatment with JH at pupariation inhibits the formation of abdominal bristles, which are derived from the histoblasts. However, the mechanism via which JH exerts this effect remains poorly understood. In this study, we analyzed the effect of JH on histoblast proliferation, migration, and differentiation. Our results indicated that whereas the proliferation and migration of histoblasts remained unaffected following treatment with a JH mimic (JHM), their differentiation, particularly the specification of sensor organ precursor (SOP) cells, was inhibited. This effect was attributable to downregulated proneural genes achaete (ac) and Scute (sc) expression levels, which prevented the specification of SOP cells in proneural clusters. Moreover, Kr-h1 was found to mediate this effect of JHM. Histoblast-specific overexpression or knockdown of Kr-h1, respectively mimicked or attenuated the effects exerted by JHM on abdominal bristle formation, SOP determination, and transcriptional regulation of ac and sc. These results indicated that the defective SOP determination was responsible for the inhibition of abdominal bristle formation by JHM, which, in turn, was mainly mediated via the transducing action of Kr-h1.

Risk Assessment of Pesticide Residues by GC-MSMS and UPLC-MSMS in Edible Vegetables.

In recent years, there has been a significant increase related to pesticide residues in foods, which may increase the risks to the consumer of these foods with the different quality and concentrations of pesticide residues. Pesticides are used for controlling pests that reduce yields. On the other hand, it has become a major public health concern due to its toxic properties. Thus, the objective of the current study employed the application of Quick Easy Cheap Effective Rugged Safe (QuEChERS) method, in combination with gas and liquid chromatography-tandem mass spectrometric detection (GCMSMS, LCMSMS) in order to determine 137 pesticide residues (63 insecticides, 41 acaricides, 40 herbicide, 55 fungicide, nematicide, growth regulator, Chitin synthesis inhibitors, and Juvenile hormone mimics), in 801 vegetables such as 139 tomatoes, 185 peppers, 217 squash, 94 eggplants, and 166 cucumbers from different locations in Hail and Riyadh cities. The results showed that the majority of pesticide residues were detected for each of the following pesticides: acetaimpride, metalaxyl, imidaclopride, bifenthrin, pyridaben, difenoconazole, and azoxystrobien, which were repeated in the samples studied 39, 21, 11, 10, 8, 7, and 5, respectively. In addition, results observed that the tomato was the most contaminated with pesticide residues; it was contaminated with 19 compounds and was followed by pepper, cucumber, and squash, and the last commodity in the contaminated ranking was eggplant. The highest calculated estimated daily intakes (EDIs) were recorded for tomatoes which were estimated between 0.013 to 0.516 mg/kg of body weight per day (bw/day) while the lowest EDIs value was between 0.000002 to 0.0005 mg/kg of bw/day for cucumber. Results indicated that the EDIs values were lower than the acceptable daily intake (ADI) values. Results observed that the most of pesticide residues exposure in food consumption in Saudi Arabia were lower than ADIs. In addition, the highest value for health risk index (HRI) was recorded with Ethion residue in tomato, but in sweet pepper, the highest value for HRI was 127.5 in the form of fipronil residue. On the other hand, results found that the highest values of HRI were 1.54, 1.61, and 0.047 for difenoconazole, bifenthrin, and pyridaben residues in squash, eggplant, and cucumber.

A new WHO bottle bioassay method to assess the susceptibility of mosquito vectors to public health insecticides: results from a WHO-coordinated multi-centre study

BackgroundThe continued spread of insecticide resistance in mosquito vectors of malaria and arboviral diseases may lead to operational failure of insecticide-based interventions if resistance is not monitored and managed efficiently. This study aimed to develop and validate a new WHO glass bottle bioassay method as an alternative to the WHO standard insecticide tube test to monitor mosquito susceptibility to new public health insecticides with particular modes of action, physical properties or both.MethodsA multi-centre study involving 21 laboratories worldwide generated data on the susceptibility of seven mosquito species (Aedes aegypti, Aedes albopictus, Anopheles gambiae sensu stricto [An. gambiae s.s.], Anopheles funestus, Anopheles stephensi, Anopheles minimus and Anopheles albimanus) to seven public health insecticides in five classes, including pyrethroids (metofluthrin, prallethrin and transfluthrin), neonicotinoids (clothianidin), pyrroles (chlorfenapyr), juvenile hormone mimics (pyriproxyfen) and butenolides (flupyradifurone), in glass bottle assays. The data were analysed using a Bayesian binomial model to determine the concentration–response curves for each insecticide–species combination and to assess the within-bioassay variability in the susceptibility endpoints, namely the concentration that kills 50% and 99% of the test population (LC50 and LC99, respectively) and the concentration that inhibits oviposition of the test population by 50% and 99% (OI50 and OI99), to measure mortality and the sterilizing effect, respectively.ResultsOverall, about 200,000 mosquitoes were tested with the new bottle bioassay, and LC50/LC99 or OI50/OI99 values were determined for all insecticides. Variation was seen between laboratories in estimates for some mosquito species–insecticide combinations, while other test results were consistent. The variation was generally greater with transfluthrin and flupyradifurone than with the other compounds tested, especially against Anopheles species. Overall, the mean within-bioassay variability in mortality and oviposition inhibition were < 10% for most mosquito species-insecticide combinations.ConclusionOur findings, based on the largest susceptibility dataset ever produced on mosquitoes, showed that the new WHO bottle bioassay is adequate for evaluating mosquito susceptibility to new and promising public health insecticides currently deployed for vector control. The datasets presented in this study have been used recently by the WHO to establish 17 new insecticide discriminating concentrations (DCs) for either Aedes spp. or Anopheles spp. The bottle bioassay and DCs can now be widely used to monitor baseline insecticide susceptibility of wild populations of vectors of malaria and Aedes-borne diseases worldwide.Graphical abstract

RNAi-mediated knockdown of juvenile hormone acid O-methyltransferase disrupts larval development in the oriental fruit fly, Bactrocera dorsalis (Hendel)

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a notoriously agricultural pest that causes serious economic losses to fruits and vegetables. Widespread insecticide resistance in B. dorsalis is a major obstacle in successful control. Therefore, new pest control strategies, such as those targeting specific genes that can block pest development, are urgently needed. In the current study, the function of JHAMT in B. dorsalis was systematically investigated. A methyltransferase gene in B. dorsalis (BdJHAMT) that is homologous to JHAMT of Drosophila melanogaster was cloned firstly. The subsequently spatiotemporal expression analysis indicated that BdJHAMT mRNA was continuously present in the larval stage, declined sharply immediately before pupation, and then increased in the adult. Subcellular localization showed that BdJHAMT was localized in the adult corpora allata and larval intestinal wall cells. The JH III titer in B. dorsalis was closely related to the transcription level of BdJHAMT in different developmental stages. The dsBdJHAMT feeding-based RNAi resulted in a greatly decreased JH III titer that disrupted fly development. The slow growth caused by BdJHAMT silencing was partially rescued by application of the JH mimic, methoprene. These results demonstrated that BdJHAMT was crucial for JH biosynthesis and thus regulated larval development in B. dorsalis, indicating it may serve as a prospective target for the development of novel control strategies against this pest.

Development and characterization of Spodoptera mauritia ovarian primary cell culture and evaluation of fenoxycarb toxicity.

The rice swarming caterpillar, Spodoptera mauritia (Biosduval, 1833) (Lepidoptera: Noctuidae), has been considered a severe pest and has caused the extreme loss of rice paddies in the nursery stages. The massive reproductive efficiency of pests results from balanced endocrine functioning. Insect treatments with exogenous either juvenile hormones (JH) or juvenile hormone analogues (JHAs) during low endogenous JH titers disrupt metamorphosis andovarian development. Hence, it was thought worthwhile to develop a primary ovarian cell culture of Spodoptera mauritia to study the biological efficiency of JHA, fenoxycarb, at the cellular level. The study envisioned the cell characteristics and growth properties and found that most cells were spherical. Spindle-shaped cells were also present during the initial stage of active cell division. The majority of the cells grew attached to the bottom of the culture plates, and a few grew in suspension. The cell doubling time was 144 ± 14h. The growth curve exhibited a sigmoid pattern of cell proliferation at first, followed by a declining phase. These properties indicate that the primary ovarian culture is a suitable candidate for developing a novel cell line of S. mauritia. The preliminary study investigated the cytotoxicity of the insect growth regulator fenoxycarb in ovarian, primary cell culture. Incubated cells showed noticeable morphological changes, including cell shrinkage and apoptotic body formation; inhibition of cellular proliferation; and apoptosis-inducing effects in ovarian, primary cell culture of S. mauritia. Evidently, from these results, it is clear that juvenile hormone mimics can react in a meaningful way to control the rice pest S. mauritia by inducing cellular apoptosis in ovarian cells, which may cause adult females to become sterile.

The impact of insect growth regulators on adult emergence inhibition and the fitness of Aedes aegypti field populations in Thailand

BackgroundThe frequent use of insecticides in vector control causes the development of insecticide resistance. Insect growth regulators (IGRs), which effect insect development, are used as a promising alternative to control resistant insect vectors. This study aimed to develop novel effective tools for Aedes aegypti control by evaluating the efficacy of different IGRs on larval development, blood feeding capacity, fecundity, and fertility in females and sperm productivity in males across geographical regions of Thailand. MethodsThe efficacy of 16 technical grade IGRs were evaluated against laboratory strain Ae. aegypti larvae in order to determine their emergence inhibition (EI) at 50% and 95% under laboratory conditions. Six IGRs were selected for fecundity, fertility, and sperm productivity studies using feed-through treatments at EI95 concentration levels against adult Ae. aegypti field strains. ResultsThe results from larval bioassay tests indicate that juvenile hormone mimics (EI50 = 0.010–0.229 ppb; EI95 = 0.066–1.118 ppb) and chitin synthesis inhibitors affecting CHS1 (EI50 = 0.240–2.412 ppb; EI95 = 0.444–4.040 ppb) groups effectively inhibited adult Ae. aegypti emergence. Methoprene and fenoxycarb significantly reduced blood feeding capacity. Egg production was comparable among strains while methoprene, pyriproxyfen and diflubenzuron induced egg production. Egg retention was detected in females fed on diflubenzuron. Methoprene, fenoxycarb, diflubenzuron, and teflubenzuron reduced egg hatching rates in mosquito field strains compared to laboratory strain. Male mosquitoes fed on fenoxycarb showed significantly lower sperm production compared to other treatments. ConclusionJuvenile hormone analogues and chitin synthesis inhibitors affecting CHS1 groups showed excellent results in adult emergence inhibition in this study. They also disrupted reproductive systems in both adult males and females. This study suggested that they can be used as an alternative larvicide in mosquito control programs.

Investigations on dynamics of juvenile hormone mimics on the economic parameters of silkworm, Bombyx mori L.

Juvenile hormone mimics are known to prolong larval life in insects and thus in sericulture, it has been shown to boost good cocoon and silk yield. In the present experiment, the double hybrid (FC1XFC2) silkworm was utilized and analysed the performance of economic traits. The influence of juvenile hormone mimics ie., phytojuvenoids from Pinus (Pinus roxburghii), Custard apple (Annona squamosa) and Tapioca (Manihot esculenta), a commercially available juvenile hormone mimic Serimore and Pyriproxyfen has been recorded as quantitative characters of silkworm when administered topically with two different doses at two distinct larval instars. The quantum of expression of economic trait like cocoon weight, shell weight, shell ratio, filament length and denier were found to be increasing with the increase in the number of larval treatments which is double administration at the rate of 20% and 30% concentration for Pinus and Tapioca treated larvae. In case of custard apple treated larvae, double treatment at 20% concentration showed best results. Single treatment at 5µl and 10µl concentration of pyriproxyfen gave similar outcome with respect to all economic parameters. Among all, Pinus was found to be on par with Serimore for all the economic characters.

Synthesis, characterization and insect growth regulating study of beta-alanine substituted sulfonamide derivatives as juvenile hormone mimics

A series of β-alanine substituted sulfonamide compounds (6a-6h) as juvenile hormone analogs (JHAs) has been synthesized. The structures of the synthesized compounds have been confirmed by FT-IR, 1H and 13C NMR spectroscopy and ESI-MS (high-resolution mass spectra [HRMS]) techniques. Biological screening of the synthesized analogs has been performed on the fifth larval instar of Galleria mellonella. JH activity of the compounds (6a-6h) has been tested at various concentrations and compared with commercially used JHAs pyriproxyfen (1), fenoxycarb (2). Compound N-methyl-N-phenyl-3-(p-tolylsulfonylamino)propanamide (6d) exhibited better activity among all the tested compounds with LC50 and LC90 values of 0.16 and 0.52 mg/mL, respectively. To understand the structure–activity relationship (SAR), quantum chemical descriptors have been calculated using density functional theory (DFT) method. Toxicity prediction has been performed using the absorption, distribution, metabolism, excretion and toxicity (ADMET) tool. The compounds are proposed to be nontoxic, eco-friendly and biodegradable.

Sexual dimorphism of diapause regulation in the hemipteran bug Pyrrhocoris apterus

Diapause is one of the major strategies for insects to prepare for and survive harsh seasons. In females, the absence of juvenile hormone (JH) is a hallmark of adult reproductive diapause, a developmental arrest, which is much less characterized in males. Here we show that juvenile hormone III skipped bisepoxide (JHSB3) titers in hemolymph remarkably differ between reproductive males and females of the linden bug Pyrrhocoris apterus, whereas no JH was detected in diapausing adults of both sexes. Like in females, ectopic application of JH mimic effectively terminated male diapause through the canonical JH receptor components, Methoprene-tolerant and Taiman. In contrast to females, long photoperiod induced reproduction even in males with silenced JH reception or in males with removed corpus allatum (CA), the JH-producing gland. JHSB3 was detected in the accessory glands (MAG) of reproductive males, unexpectedly, even in males without CA. If there is a source of JHSB3 outside CA or a long-term storage of JHSB3 in MAGs remains to be elucidated. These sex-related idiosyncrasies are further manifested in different dynamics of diapause termination in P. apterus by low temperature. We would like to propose that this sexual dimorphism of diapause regulation might be explained by the different reproductive costs for each sex.

Binding of de novo synthesized radiolabeled juvenile hormone (JH III) by JH receptors from the Cuban subterranean termite Prorhinotermes simplex and the German cockroach Blattella germanica

Juvenile hormone (JH) controls insect reproduction and development through an intracellular receptor complex comprising two bHLH-PAS proteins, the JH-binding Methoprene-tolerant (Met) and its partner Taiman (Tai). Many hemimetabolous insects including cockroaches strictly depend on JH for stimulation of vitellogenesis. In termites, the eusocial hemimetabolans, JH also regulates the development of caste polyphenism. Studies addressing the agonist ligand binding to recombinant JH receptors currently include three species belonging to two holometabolous insect orders, but none that would represent any of the hemimetabolous orders. Here, we examined JH receptors in two representatives of Blattodea, the cockroach Blattella germanica and the termite Prorhinotermes simplex. To test the JH-binding capacity of Met proteins from these species, we performed chemical synthesis and tritium labeling of the natural blattodean JH homolog, JH III. Our improved protocol increased the yield and specific activity of [10-3H]JH III relative to formerly available preparations. Met proteins from both species specifically bound [3H]JH III with high affinity, whereas Met variants mutated at a critical position within the ligand-binding domain were incapable of such binding. Furthermore, JH III and the synthetic JH mimic fenoxycarb stimulated dimerization between Met and Tai components of the respective JH receptors of both species. These data present primary evidence for agonist binding by JH receptors in any hemimetabolous species and provide a molecular basis for JH action in cockroaches and termites.

Synthesis, structural, DFT and bio-assay of novel ethyl 3-(2-(4-chlorophenoxy)acetamido) propanoate on Galleria mellonela (wax moth) – a juvenile hormone mimic as potential insect growth regulator (IGR)

Ethyl 3-(2-(4-chlorophenoxy)acetamido)propanoate is synthesized and comprehensively studied as an insect growth regulator. The structure has been confirmed using fourier transform infra-red (FT-IR), nuclear magnetic resonance (1D-NMR, 2D-NMR), electrospray ionization (ESI-MS) spectroscopic techniques. The synthesized compound has been tested against the fifth instar of Galleria mellonella in comparison to pyriproxyfen (commercial IGR). Observed LC50 and LC90 values of synthesized compound and pyriproxyfen are 162 ppm & 549 ppm and 125 & 630 ppm respectively. Computational and structural parameters of the synthesized compound are determined and studied. Molecular geometry and unscaled vibrational frequencies are calculated by density functional theory (DFT) method using Becke's three-parameter lee-yang-parr (B3LYP) hybrid functional at 6–311G+(d,2p) level of basis set. The scaled quantum mechanical (SQM) method has been employed to calculate scaled vibrational frequencies. Further, 1H and 13C NMR spectra are calculated with (DFT/ B3LYP) using gage independent atomic orbital method (GIAO) in the gaseous and solvent phases to confirm the structure of synthesized compound. Natural bond orbital (NBO) analysis has been used to study the intramolecular charge transfer (ICT) interactions, delocalization of electron density. HOMO-LUMO energies are calculated to describe global reactivity descriptors which played an important role in explaining the bioactivity of compounds. Parameters are also evaluated in solvent phases using the integral equation formalism polarization continuum model (IEF-PCM) of the DFT method. The Autodock software tool is used to study the binding interactions between ligand and receptor juvenile hormone binding protein (JHBP).

Statewide Efficacy Assessment of Insect Growth Regulators Against Aedes albopictus (Diptera: Culicidae) in Sabah, Malaysia: An Alternative Control Strategy?

The efficacy of three groups of insect growth regulators, namely juvenile hormone mimics (methoprene and pyriproxyfen), chitin synthesis inhibitors (diflubenzuron and novaluron), and molting disruptor (cyromazine) was evaluated for the first time, against Aedes albopictus Skuse (Diptera: Culicidae) larvae from 14 districts in Sabah, Malaysia. The results showed that all field populations of Ae. albopictus were susceptible towards methoprene, pyriproxyfen, diflubenzuron, novaluron, and cyromazine, with resistance ratio values ranging from 0.50-0.90, 0.60-1.00, 0.67-1.17, 0.71-1.29, and 0.74-1.07, respectively. Overall, the efficacy assessment of insect growth regulators in this study showed promising outcomes and they could be further explored as an alternative to conventional insecticides.

Histone H3K27 methylation–mediated repression of Hairy regulates insect developmental transition by modulating ecdysone biosynthesis

Insect development is cooperatively orchestrated by the steroid hormone ecdysone and juvenile hormone (JH). The polycomb repressive complex 2 (PRC2)-mediated histone H3K27 trimethylation (H3K27me3) epigenetically silences gene transcription and is essential for a range of biological processes, but the functions of H3K27 methylation in insect hormone action are poorly understood. Here, we demonstrate that H3K27 methylation-mediated repression of Hairy transcription in the larval prothoracic gland (PG) is required for ecdysone biosynthesis in Bombyx and Drosophila H3K27me3 levels in the PG are dynamically increased during the last larval instar. H3K27me3 reduction induced by the down-regulation of PRC2 activity via inhibitor treatment in Bombyx or PG-specific knockdown of the PRC2 component Su(z)12 in Drosophila diminishes ecdysone biosynthesis and disturbs the larval-pupal transition. Mechanistically, H3K27 methylation targets the JH signal transducer Hairy to repress its transcription in the PG; PG-specific knockdown or overexpression of the Hairy gene disrupts ecdysone biosynthesis and developmental transition; and developmental defects caused by PG-specific Su(z)12 knockdown can be partially rescued by Hairy down-regulation. The application of JH mimic to the PG decreases both H3K27me3 levels and Su(z)12 expression. Altogether, our study reveals that PRC2-mediated H3K27 methylation at Hairy in the PG during the larval period is required for ecdysone biosynthesis and the larval-pupal transition and provides insights into epigenetic regulation of the crosstalk between JH and ecdysone during insect development.

Characterization of E93 in neometabolous thrips Frankliniella occidentalis and Haplothrips brevitubus.

Insect metamorphosis into an adult occurs after the juvenile hormone (JH) titer decreases at the end of the juvenile stage. This generally coincides with decreased transcript levels of JH-response transcription factors Krüppel homolog 1 (Kr-h1) and broad (br), and increased transcript levels of the adult specifier E93. Thrips (Thysanoptera) develop through inactive and non-feeding stages referred to as “propupa” and “pupa”, and this type of distinctive metamorphosis is called neometaboly. To understand the mechanisms of hormonal regulation in thrips metamorphosis, we previously analyzed the transcript levels of Kr-h1 and br in two thrips species, Frankliniella occidentalis (Thripidae) and Haplothrips brevitubus (Phlaeothripidae). In both species, the transcript levels of Kr-h1 and br decreased in the “propupal” and “pupal” stages, and their transcription was upregulated by exogenous JH mimic treatment. Here we analyzed the developmental profiles of E93 in these two thrips species. Quantitative RT-PCR revealed that E93 expression started to increase at the end of the larval stage in F. occidentalis and in the “propupal” stage of H. brevitubus, as Kr-h1 and br mRNA levels decreased. Treatment with an exogenous JH mimic at the onset of metamorphosis prevented pupal-adult transition and caused repression of E93. These results indicated that E93 is involved in adult differentiation after JH titer decreases at the end of the larval stage of thrips. By comparing the expression profiles of Kr-h1, br, and E93 among insect species, we propose that the “propupal” and “pupal” stages of thrips have some similarities with the holometabolous prepupal and pupal stages, respectively.

Effects of fragrance compounds on growth of the silkworm Bombyx mori.

Due to the contamination and biological toxicity of some fragrance compounds, the environmental and ecological problems of such compounds have attracted more and more attention. However, studies of the toxicity of fragrance compounds for insects have been limited. The toxicity of 48 fragrance compounds for the silkworm Bombyx mori were investigated in this study. All of the fragrance compounds examined had no acute toxicity for B. mori larvae, but eight of them (menthol, maltol, musk xylene, musk tibeten, dibutyl sulfide, nerolidol, ethyl vanillin, and α-amylcinnamaldehyde) exhibited chronic and lethal toxicity with LC50 values from 20 to 120 µM. In a long-term feeding study, musk tibeten, nerolidol, and musk xylene showed significant growth regulatory activity. They were also extremely harmful to the cocooning of B. mori, resulting in small, thin, and loose cocoons. Two important insect hormones, namely, juvenile hormone (JH) and 20-hydroxyecdysone (20-E), were quantified in hemolymph following chronic exposure to musk tibeten, nerolidol, and musk xylene, respectively. Musk tibeten significantly increased JH titer and decreased the 20-E titer in hemolymph, and musk xylene had a significant inhibitory effect on JH titer and increased 20-E titer. Although nerolidol had no effect on hormone levels, exogenous JH mimic nerolidol increased the physiological effects of JH and significantly slowed the growth rate of B. mori larvae. The results showed that these fragrance compounds could interfere with the insect endocrine system, leading to death and abnormal growth. The risk to insects of residual fragrance compounds in the environment is worthy of attention.

Ovicidal activity of juvenile hormone mimics in the bean bug, Riptortus pedestris.

Insect juvenile hormone (JH) mimics (JHMs) are known to have ovicidal effects if applied to adult females or eggs. Here, we examined the effects of exogenous JHMs on embryonic development of the bean bug, Riptortus pedestris. The expression profiles of JH early response genes and JH biosynthetic enzymes indicated that JH titer was low for the first 3 days of the egg stage and increased thereafter. Application of JH III skipped bisepoxide (JHSB3) or JHM on Day 0 eggs when JH titer was low caused reduced hatchability, and the embryos mainly arrested in mid- or late embryonic stage. Application of JHMs on Day 5 eggs also resulted in an arrest, but this was less effective compared with Day 0 treatment. Interestingly, ovicidal activity of synthetic JHMs was much lower than that of JHSB3. This study will contribute to developing novel insecticides that are selective among insect species.

Lethal and Sublethal Effects of Pyriproxyfen on Apis and Non-Apis Bees.

Pyriproxyfen is a juvenile hormone mimic used extensively worldwide to fight pests in agriculture and horticulture. It also has numerous applications as larvicide in vector control. The molecule disrupts metamorphosis and adult emergence in the target insects. The same types of adverse effects are expected on non-target insects. In this context, the objective of this study was to evaluate the existing information on the toxicity of pyriproxyfen on the honey bee (Apis mellifera) and non-Apis bees (bumble bees, solitary bees, and stingless bees). The goal was also to identify the gaps necessary to fill. Thus, whereas the acute and sublethal toxicity of pyriproxyfen against A. mellifera is well-documented, the information is almost lacking for the non-Apis bees. The direct and indirect routes of exposure of the non-Apis bees to pyriproxyfen also need to be identified and quantified. More generally, the impacts of pyriproxyfen on the reproductive success of the different bee species have to be evaluated as well as the potential adverse effects of its metabolites.

Deployment and Fact Analysis of the In2Care® Mosquito Trap, A Novel Tool for Controlling Invasive Aedes Species.

During April-October 2019, the West Valley Mosquito and Vector Control District (Ontario, CA) deployed large numbers of In2Care® mosquito traps in a preliminary study to evaluate the trap's potential effectiveness at controlling invasive Aedes aegypti (L.) and Ae. albopictus (Skuse) in 6 cities of San Bernardino County, CA. The trap was used to attract ovipositing females, expose them to the juvenile hormone mimic pyriproxyfen and the entomopathogenic fungus Beauveria bassiana, and autodisseminate pyriproxyfen to other water sources prior to their death from fungal infection. The trap attracted Ae. aegypti and Culex quinquefasciatus, with the latter species predominating at much higher larval densities in the trap reservoirs. Field-collected larvae and pupae from the trap reservoirs showed complete adult emergence inhibition. Furthermore, the trap reservoirs retained high levels of residual larvicidal, pupicidal, and emergence inhibition activity after they were retrieved from the field, as indicated by laboratory bioassays against laboratory colony of Cx. quinquefasciatus. Results of this study support more detailed quantitative local evaluations on trap efficacy to measure the impact of the In2Care mosquito trap on wild invasive Aedes and Culex populations in future mosquito control efforts.

Involvement of the transcription factor E75 in adult cuticular formation in the red flour beetle Tribolium castaneum

Insect adult metamorphosis generally proceeds with undetectable levels of juvenile hormone (JH). In adult development of the red flour beetle Tribolium castaneum, biosynthesis of adult cuticle followed by its pigmentation and sclerotization occurs, and dark coloration of the cuticle becomes visible in pharate adults. Here, we examined the molecular mechanism of adult cuticular formation in more detail. We noticed that an exogenous JH mimic (JHM) treatment of Day 0 pupae did not inhibit pigmentation or sclerotization, but instead, induced precocious pigmentation of adult cuticle two days in advance. Quantitative RT-PCR analyses revealed that ecdysone-induced protein 75B (E75) is downregulated in JHM-treated pupae. Meanwhile, tyrosine hydroxylase (Th), an enzyme involved in cuticular pigmentation and sclerotization, was precociously induced, whereas a structural cuticular protein CPR27 was downregulated, by exogenous JHM treatment. RNA interference-mediated knockdown of E75 resulted in precocious adult cuticular pigmentation, which resembled the phenotype caused by JHM treatment. Notably, upregulation of Th as well as suppression of CPR27 were observed with E75 knockdown. Meanwhile, JHM treatment suppressed the expression of genes involved in melanin synthesis, such as Yellow-y and Laccase 2, but E75 knockdown did not result in marked reduction in their expression. Taken together, these results provided insights into the regulatory mechanisms of adult cuticular formation; the transcription of genes involved in adult cuticular formation proceeds in a proper timing with undetectable JH, and exogenous JHM treatment disturbs their transcription. For some of these genes such as Th and CPR27, E75 is involved in transcriptional regulation. This study shed light on the molecular mode of action of JHM as insecticides; exogenous JHM treatment disturbed the expression of genes involved in the adult cuticular formation, which resulted in lethality as pharate adults.

Diminished fitness of red cotton bug Dysdercus koenigii Fabricius induced by the volatiles in the hexane extract of Catharanthus roseus

Survival, longevity, growth index and development of fifth instar nymphs of Dysdercus koenigii (Heteroptera: Pyrrhocoriedae) as influenced by the Catharanthus roseus was assessed in the present investigation. The fifth instar nymphs were exposed to the volatiles present in the extract for 24 h. Different concentrations of the extract ranging from 0.00625% to 10% were used for the treatment. The results indicate that the survival of treated nymphs was not affected significantly. However, the volatiles present in the extract influenced the growth and development of the treated fifth instar nymphs. The nymphs exhibited decreased growth as reflected by the decrease in growth index. The extract induced different types of developmental anomalies notably incomplete moulting, supernumerary instars, adultoids and adults with deformed wing. In some cases, exuviae did not detach from the body of the adults and create obstacle in the feeding, walking and mating. The effects of the extract on nymphal development were also dose-dependent. No normal adults could develop in the treatments with the concentration 5% or more; various morphological and developmental abnormalities were reported in the nymphs treated with 2.5% to 0.1% of the extract. GC-MS analysis of the hexane leaf extract of C. roseus showed the presence of sesquiterpenes including geranylgerniol and many toxic chemicals. These phytoconstituents of the C. roseus extract besides having cidal effects also showed juvenile hormone mimic activities which impair lifetime fitness of D. koenigii. Therefore, botanical from the C. roseus can potentially be used as an alternative approach in pest management.